The overall vision of the proposed research is to create an effective, scalable, and affordable biomimetic corneal substitute suitable for clinical translation. Damage to the cornea, the outermost tissue of the eye, causes loss of vision, which profoundly impacts quality of life. Cornea transplantation is the gold standard for treatment, but slowly progressing rejection, short supply of donor tissue, and the small number of failures that lead to devastating consequences, remain key challenges. Artificial corneal substitutes have thus far failed to recapitulate the native tissue structure and function. An effective biosynthetic replacement that mimics the native cornea would offer a transformational new option for corneal injury patients. Our prior work has opened a novel route to fully functional synthetic corneas. We previously developed a collagen vitrification process that increases the mechanical strength of collagen gels. To achieve the necessary transparency in full thickness implants, we needed to further control collagen spacing and lamellae orthogonality. We recapitulated cornea development and organization in vitro using synthetic cyclic proteoglycan molecules, cyclodextrins (CD). Preliminary data with CD and collagen vitrification produced materials with aligned fibrillar architecture and orthogonally organized lamellae similar to the native cornea. Furthermore, cells were able to grow on the materials and implants were suturable and biocompatible in a rabbit corneal defect. Our objective is to replicate the native cornea ultrastructure, including structure (collagen fibril size, alignment and lamella) and function (transparency and mechanical strength). We hypothesize that the CD molecules interact specifically with collagen to control fiber spacing and assembly during vitrification, enabling a fully functional corneal replacement material. To achieve these goals, we propose the following specific aims: Specific Aim 1: Develop a library of CD-Collagen biomaterials using statistical optimization. A CD-Col material library will be synthesized with varying forms of ?, ?, and ? CDs using multiphase statistical optimization Design of Experiments (DOE) will be used to define and optimize corneal biomimetic structure. Specific Aim 2: Characterize physical properties of CD-Collagen biomaterials. Formulations from Aim 1 that achieve minimum clarity and mechanics for corneal implantation will be characterized and compared to the native cornea and standard collagen implants. Specific Aim 3: Translate the biomimetic CD-Collagen material to a corneal defect model.